Abstract
Hepatocytes comprise the majority of liver mass and cell number. However, in order to understand liver biology, the non-parenchymal cells (NPCs) must be considered. Herein, a relatively rapid and efficient method for isolating liver NPCs from a mouse is described. Using this method, liver sinusoidal endothelial cells, Kupffer cells, natural killer (NK) and NK-T cells, dendritic cells, CD4+ and CD8+ T cells, and quiescent hepatic stellate cells can be purified. This protocol permits the collection of peripheral blood, intact liver tissue, and hepatocytes, in addition to NPCs. In situ perfusion via the portal vein leads to efficient liver digestion. NPCs are enriched from the resulting single-cell suspension by differential and gradient centrifugation. The NPCs can by analyzed or sorted into highly enriched populations using flow cytometry. The isolated cells are suitable for flow cytometry, protein, and mRNA analyses as well as primary culture.
Isaac Mohar and Katherine J. Brempelis are co-first authors of this chapter.
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Acknowledgements
This protocol was developed under support by the National Institutes of Health grants R21AI099872 and T32AI007411 and Seattle Biomedical Research Institute.
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Mohar, I., Brempelis, K.J., Murray, S.A., Ebrahimkhani, M.R., Crispe, I.N. (2015). Isolation of Non-parenchymal Cells from the Mouse Liver. In: Vaughan, A. (eds) Malaria Vaccines. Methods in Molecular Biology, vol 1325. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2815-6_1
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DOI: https://doi.org/10.1007/978-1-4939-2815-6_1
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2814-9
Online ISBN: 978-1-4939-2815-6
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